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January 2023; 10 (1) Research ArticleOpen Access

CD59 Expression in Skeletal Muscles and Its Role in Myasthenia Gravis

Kazuo Iwasa, Yutaka Furukawa, Hiroaki Yoshikawa, Masahito Yamada, Kenjiro Ono
First published November 17, 2022, DOI: https://doi.org/10.1212/NXI.0000000000200057
Kazuo Iwasa
From the Department of Health and Medical Sciences (K.I.), Ishikawa Prefectural Nursing University; Department of Neurology (K.I., Y.F., M.Y., K.O.), Kanazawa University Graduate School of Medical Science; Department of Neurology (Y.F.), National Hospital Organization Kanazawa Medical Center; Health Service Center (H.Y.), Kanazawa University; and Department of Neurology (M.Y.), Kudanzaka Hospital.
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Yutaka Furukawa
From the Department of Health and Medical Sciences (K.I.), Ishikawa Prefectural Nursing University; Department of Neurology (K.I., Y.F., M.Y., K.O.), Kanazawa University Graduate School of Medical Science; Department of Neurology (Y.F.), National Hospital Organization Kanazawa Medical Center; Health Service Center (H.Y.), Kanazawa University; and Department of Neurology (M.Y.), Kudanzaka Hospital.
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Hiroaki Yoshikawa
From the Department of Health and Medical Sciences (K.I.), Ishikawa Prefectural Nursing University; Department of Neurology (K.I., Y.F., M.Y., K.O.), Kanazawa University Graduate School of Medical Science; Department of Neurology (Y.F.), National Hospital Organization Kanazawa Medical Center; Health Service Center (H.Y.), Kanazawa University; and Department of Neurology (M.Y.), Kudanzaka Hospital.
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Masahito Yamada
From the Department of Health and Medical Sciences (K.I.), Ishikawa Prefectural Nursing University; Department of Neurology (K.I., Y.F., M.Y., K.O.), Kanazawa University Graduate School of Medical Science; Department of Neurology (Y.F.), National Hospital Organization Kanazawa Medical Center; Health Service Center (H.Y.), Kanazawa University; and Department of Neurology (M.Y.), Kudanzaka Hospital.
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Kenjiro Ono
From the Department of Health and Medical Sciences (K.I.), Ishikawa Prefectural Nursing University; Department of Neurology (K.I., Y.F., M.Y., K.O.), Kanazawa University Graduate School of Medical Science; Department of Neurology (Y.F.), National Hospital Organization Kanazawa Medical Center; Health Service Center (H.Y.), Kanazawa University; and Department of Neurology (M.Y.), Kudanzaka Hospital.
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Citation
CD59 Expression in Skeletal Muscles and Its Role in Myasthenia Gravis
Kazuo Iwasa, Yutaka Furukawa, Hiroaki Yoshikawa, Masahito Yamada, Kenjiro Ono
Neurol Neuroimmunol Neuroinflamm Jan 2023, 10 (1) e200057; DOI: 10.1212/NXI.0000000000200057

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    Figure 1 Immunohistochemistry of the Coexpression of CD59 and Acetylcholine Receptors in Muscles

    (A) Immunohistochemistry in muscles for CD59 (green), α-BTx (red), and cell nuclei (blue), and the merging of the 3 pictures. Patient 6 with MG displays a weakly concentrated CD59 expression at the destructive α-BTx staining region (arrowhead). Patient 11 displays a weakly concentrated CD59 expression with the α-BTx staining (arrowhead), and an extra localized CD59 expression without an α-BTx staining is confirmed (arrows). Patients 9 and 10 display coexpression of CD59 and α-BTx, and a strongly localized CD59 expression is recognized near the α-BTx staining region (arrows). Patient 13 displays a CD59 localized region and a partially overlapped α-BTx staining region. Patient 16 displays a distinct coexpression of CD59 and α-BTx, similar to the nonmyopathy controls. Scale bar = 25 µm. (B) Western blotting for the detection of CD59 in nonmyopathy controls and patients with MG. Lanes 1–3 depict nonmyopathy controls, where CD59 protein is detected as weak bands. Lanes 4–8 depict patients with MG, where CD59 protein bands vary among the patients. CD59 overexpression was confirmed in 7 of 16 patients with MG. AChR = acetylcholine receptor; CD59 = cluster of differentiation 59; MG = myasthenia gravis; NMJ = neuromuscular junction; α-BTx = α-bungarotoxin.

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    Figure 2 Expressions of CD59 and Acetylcholine Receptors in Muscles of Patients With MG

    (A) Scatter plots of the quantification by western blotting for CD59, in relation to GAPDH, in nonmyopathy controls and patients with MG. The ratio of CD59/GAPDH in patients with MG was significantly higher compared with that in the nonmyopathy controls. (B) Fold changes of CD59 mRNA in nonmyopathy controls and patients with MG. The CD59 mRNA expression showed no significant difference between the nonmyopathy controls and the patients with MG (MG; n = 16, median 0.39, IQR 0.28–1.43 and nonmyopathy controls; n = 6, median 0.28, IQR 0.10–0.60, p = 0.231). (C) Fold changes of AChR mRNA in nonmyopathy controls and patients with MG. The AChR mRNA expression showed no significant difference between the nonmyopathy controls and the patients with MG. AChR mRNA expression could not be detected by qRT-PCR using TaqMan probes for CHRNA1 (Hs00175578_m1) in 1 patient with MG (MG; n = 15, median 1.08, IQR 0.40–1.87 and nonmyopathy controls; n = 6, median 0.55, IQR 0.18–0.83, p = 0.095). AChR = acetylcholine receptor; CD59 = cluster of differentiation 59; GAPDH = glyceraldehyde-3-phosphate dehydrogenase; IQR = interquartile range; MG = myasthenia gravis.

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    Figure 3 Correlation Between CD59 Expression and Clinical Scores of MG

    (A) Correlation between the fold changes in CD59 mRNA and the protein ratio of CD59/GAPDH by western blotting in nonmyopathy controls and patients with MG. Pearson correlation was used to calculate the association. (B) Correlation between the fold changes in CD59 mRNA and AChR mRNA in nonmyopathy controls and patients with MG. Pearson correlation was used to calculate the association. (C) Association between ΔCtCD59/AChR (= AChR Ct − CD59 Ct) and QMG scores in patients with MG. Pearson correlation was used to calculate the association. (D) Association between ΔCtCD59/AChR (= AChR Ct − CD59 Ct) and MG-ADL scores in patients with MG. Pearson correlation was used to calculate the association. (E) Association between ΔCtCD59/AChR (= AChR Ct − CD59 Ct) and MGFA Clinical Classification in patients with MG. Spearman correlation was used to calculate the association. AChR = acetylcholine receptor; CD59 = cluster of differentiation 59; GAPDH = glyceraldehyde-3-phosphate dehydrogenase; MG = myasthenia gravis; MG-ADL = MG-specific activities of daily living; MGFA = Myasthenia Gravis of Foundation of America; QMG = quantitative MG.

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